Defining the roles of an enhancer long non-coding RNA eIncRNA-ID2 in rickettsial pathogenesis and immunity

定义增强子长链非编码 RNA eIncRNA-ID2 在立克次体发病机制和免疫中的作用

• 批准号: 10323675
• 负责人: Abha Sahni
• 金额: $ 19.75万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-04 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10323675
• 关键词: Address; Affect; Agreement; Animal Model; Anti-Infective Agents; Area; B-Lymphocytes; Bacteria; Biogenesis; Biological; Biological Process; Biology; Blood; Brain; C3H/HeN Mouse; CD8-Positive T-Lymphocytes; Cells; Cellular Immunity; Cerebral Edema; ChIP-seq; Chemosensitization; Code; Communicable Diseases; DNA; Data Set; Dendritic Cells; Deoxyribonuclease I; Development; Disease; Disease Progression; E protein; EP300 gene; Edema; Elements; Encyclopedia of DNA Elements; Endothelial Cells; Enhancers; Epigenetic Process; Functional disorder; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic Transcription; Genomics; Goals; Helix-Loop-Helix Motifs; Human; Human Genome; Hypersensitivity; ID2 gene; Immune; Immune System Diseases; Immune response; Immune system; Immunity; Immunology; In Vitro; Infection; Inflammation; Inhibitor of Differentiation Proteins; Investigation; Knowledge; Link; Lung; Lysine; Maintenance; Mediating; Messenger RNA; Microbiology; Modification; Molecular; Mus; Natural Killer Cells; Nucleotides; Organ; Outcome; POLR2A gene; Pathogenesis; Pathogenicity; Physiological; Poly(A) Tail; Predisposition; Proteins; Publishing; RNA; RNA Polymerase II; RNA Splicing; Regulation; Regulator Genes; Rickettsia; Rickettsia Infections; Rickettsia conorii; Role; Shapes; Site; Spleen; System; T memory cell; T-Lymphocyte; Testing; Therapeutic; Tissues; Transcript; Transcription Initiation Site; Transcriptional Regulation; Untranslated RNA; Up-Regulation; Vaccine Adjuvant; Vascular Permeabilities; alpha helix; antagonist; base; body system; cell mediated immune response; cell type; dark matter; defined contribution; follow-up; histone methylation; human disease; human model; immune activation; in vivo Model; insight; macrophage; mammalian genome; microbial; migration; monolayer; mouse model; novel; novel strategies; pathogenic microbe; polypeptide; prevent; programs; promoter; response; spotted fever; therapeutic development; therapeutic target; tick-borne; transcription factor; transcriptome sequencing; vascular inflammation

PROJECT SUMMARY Precise and dynamic alterations in gene expression are critical determinants of the regulation of host immunity to microbial pathogens. Pathogenic rickettsiae in the spotted fever group cause some of the most severe infectious diseases in humans, characterized by microvascular inflammation and dysfunction attributed to disseminated infection of endothelial cells and increased vascular permeability resulting in pulmonary/cerebral edema. Long non-coding (lnc) RNAs of ≥ 200 nucleotides regulate a panoply of biological responses through an array of mechanisms and changes in their expression levels are now intricately linked to the determination of innate as well as cell-mediated immune responses. As an important subset of lncRNAs, enhancer lncRNAs implement their regulatory roles by enhancing protein coding genes (PCGs) in a cis- or trans-acting manner. We performed RNA-sequencing on the lungs as one of the predominantly affected target organs of susceptible mice infected with R. conorii to identify up-regulation of 179 lncRNAs. Via follow-up analysis to differentiate enhancer (elnc) from promoter-associated (plnc) RNAs based on the ratio of single- versus tri-methylation of histone 3 at lysine 4 (H3K4Me1:H3K4Me3) and other active enhancer signatures based on POLR2A, p300, DNase I hypersensitivity sites, CTCF, and Hi-3C ChIP-Seq datasets, we further determined significantly higher expression of an active elncRNA013718 and its target PCG Inhibitor of DNA binding 2 (ID2) in the mouse lungs, spleen, and CD8+ T-cells during Rickettsia conorii infection. Our preliminary findings further suggest that elncRNA013718 positively regulates the expression of ID2, a protein antagonist of E protein transcription factors and a regulator of T cells in the immune system. Accordingly, we refer to elncRNA013718 as elncRNA- ID2 and hypothesize novel contributory roles for elncRNA-ID2:Id2 interplay in the regulation of protective host immunity during rickettsial infections. We propose to test this hypothesis via two independent yet thematically interlinked specific aims. Aim 1 will distinguish cell type-specific expression and functional roles of elncRNA- ID2 and ID2 in the host lungs and spleen in experimental murine models of R. conorii and R. australis infection. In Aim 2, we will determine the modulatory effects of both global and cell-specific interference with elncRNA- ID2 on host immune responses and disease progression/outcome. Given the complexity of cellular immune responses, we will employ two independent and established in vivo models of infection closely mimicking the pathophysiology of human rickettsioses and cutting-edge approaches of cellular microbiology and immunology to determine the regulatory potential of elncRNA-ID2 as a novel elncRNA in the molecular circuitry underlying regulation of host immunity. The acquired insights will enhance our knowledge of context-specific physiological roles of elncRNA-ID2 in the determination of host responses to pathogenic rickettsiae and reveal potentially unique entry points for novel strategies to enhance host immunity against intracellular microbial infections.

项目摘要 基因表达的精确和动态变化是宿主免疫调节的关键决定者 到微生物病原体。斑点发烧组中的致病性人力体造成了一些最严重的 人类感染性疾病的特征是微血管感染和功能障碍归因于 内皮细胞的传播感染并增加血管通透性，导致肺部/脑部 浮肿。 ≥200个核苷酸的长非编码（LNC）RNA通过 现在，一系列机制及其表达水平的变化与确定相关 先天和细胞介导的免疫复杂。作为LNCRNA的重要子集，增强子lncrnas 通过以顺式或反式作用方式增强蛋白质编码基因（PCG）来实现其调节作用。 我们在肺部进行了RNA测序，作为易感的主要影响的目标器官之一 感染了康罗伊菌的小鼠以鉴定179 lncRNA的上调。通过后续分析来区分 基于启动子相关（PLNC）RNA的增强子（ELNC），基于单与三甲基的比率 赖氨酸4（H3K4ME1：H3K4ME3）的Hisstone 3和其他基于Polr2A的活跃增强剂签名，P300， DNase I超敏反应位点，CTCF和HI-3C芯片seq数据集，我们进一步确定了更高的确定 活性ElncRNA013718及其靶PCG抑制剂的DNA结合2（ID2）的表达 立克康罗伊感染期间的肺，脾和CD8+ T细胞。我们的初步发现进一步表明 ELNCRNA013718积极调节ID2的表达，ID2是E蛋白转录的蛋白拮抗剂 免疫系统中T细胞的因素和调节剂。彼此之间，我们将elncrna013718称为elncrna- ID2和假设的ElncRNA-ID2：ID2相互作用的新型贡献作用在保护宿主的调节中 立克感染期间的免疫力。我们建议通过两个独立但主题来检验这一假设 相互联系的特定目标。 AIM 1将区分细胞类型特异性表达和elncrna-的功能作用 宿主肺和脾脏中的ID2和ID2在R. conorii和R. australis感染的实验鼠模型中。 在AIM 2中，我们将确定全球和细胞特异性干扰对Elncrna-的调节作用 宿主免疫调查和疾病进展/结果的ID2。鉴于细胞免疫复杂的复杂性 回答，我们将采用两个独立并在体内建立的感染模型，以密切模仿 人体力学的病理生理学和细胞微生物学和免疫学的尖端方法 确定elncrna-id2作为分子电路中新型elncrna的调节势 宿主免疫的调节。获得的见解将增强我们对特定环境生理的知识 elncrna-id2在确定宿主对病原体立克的反应中的作用，并可能揭示了潜在的 新型策略的独特入口点，以增强对细胞内微生物感染的免疫力。